title: Developmental competence of oocytes isolated from surplus medulla tissue in connection with cryopreservation of ovarian tissue for fertility preservation.
abstract: OBJECTIVE
  Evaluating the developmental competence of immature oocytes collected from surplus medulla tissue in connection with ovarian tissue cryopreservation for fertility preservation.
  
  
  METHODS
  Cohort comparative study.
  
  
  METHODS
  University laboratory in Denmark from 2011-2012.
  
  
  METHODS
  69 girls and women (0-38 years of age) who each had one ovary cryopreserved for fertility preservation.
  
  
  METHODS
  Ovaries were obtained directly from the local hospital or from collaborating hospitals (two to five hours' transport on ice). Immature oocytes were aspirated from large antral follicles visible on the ovaries, and collected from the saline solution, containing surplus medulla tissue, following dissection of the ovarian cortical tissue for cryopreservation. The immature oocytes were cultured for 48 h in an Embryoscope™ Time-lapse System or in culture dishes overlaid with liquid paraffin using commercial and in-house supplemented culture media.
  
  
  METHODS
  Maturation rate for immature oocytes reaching metaphase II.
  
  
  RESULTS
  With a maturation rate of 3.1%, only 21 of 682 immature oocytes reached metaphase II. Immature oocytes from ovaries that had been transported on ice for two to five hours performed significantly poorer than those recovered immediately after surgery. Addition of epidermal growth factor and follicle fluid from human small antral follicles to the culture medium did not augment the maturation rate. Immature oocytes cultured in the Embryoscope performed significantly better than those in conventional culture dishes.
  
  
  CONCLUSIONS
  In vitro maturation of immature oocytes should only be attempted clinically from visible antral follicles and where the ovary is not subjected to a cooling period prior to recovery of immature oocytes.
authors.affiliation: Laboratory of Reproductive Biology, Juliane Marie Center for Women, Children and Reproduction, University Hospital of Copenhagen, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Helle N
authors.initials: HN
authors.lastname: Wilken-Jensen
authors.forename: Stine G
authors.initials: SG
authors.lastname: Kristensen
authors.forename: Janni V
authors.initials: JV
authors.lastname: Jeppesen
authors.forename: Claus
authors.initials: C
authors.lastname: Yding Andersen
mesh_terms: D000293:Adolescent; D000328:Adult; D002648:Child; D002675:Child, Preschool; D015925:Cryopreservation / Q000379:methods*; D005260:Female; D059247:Fertility Preservation / Q000379:methods*; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D009865:Oocytes / Q000166:cytology / Q000254:growth & development*; D006080:Ovarian Follicle / Q000166:cytology / Q000254:growth & development*
keywords: Immaturity; embryoscope; epidermal growth factor; fertility preservation; in-vitro maturation; oocytes